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Fluorescent probes along with fluorescence microscopy are essential tools for biomedical research. Various cellular ubiquitous chemical factors such as pH, H2O2, and Ca2+ are labeled and traced using specific fluorescent probes, therefore helping us to explore their physiological function and pathological change. Among them, intracellular pH value is an important factor that governs biological processes, generally ∼7.2. Furthermore, specific organelles within cells possess unique acid-base homeostasis, involving the acidic lysosomes, alkalescent mitochondria, and neutral endoplasmic reticulum and Golgi apparatus, which undergo various physiological processes such as intracellular digestion, ATP production, and protein folding and processing. In this review, recently reported fluorescent probes targeted toward the lysosomes, mitochondria, endoplasmic reticulum, Golgi apparatus, and cytoplasm for sensing pH change are discussed, which involves molecular structures, fluorescence behavior, and biological applications.

We report herein a new class of synthetic reagents for targeting the element for nuclear expression (ENE) in MALAT1, a long noncoding RNA upregulated in many cancers. The cis-acting ENE contains a U-rich internal loop (URIL) that forms an 11 base UAU-rich triplex stem with the truncated 3' oligo-A tail of MALAT1, protecting the terminus from exonuclease digestion and greatly extending transcript lifetime. Bifacial peptide nucleic acids (bPNAs) similarly bind URILs via base triple formation between two uracil bases and a synthetic base, melamine. We synthesized a set of low molecular weight bPNAs composed of α-linked peptide, isodipeptide, and diketopiperazine backbones and evaluated their ENE binding efficacy in vitro via oligo-A strand displacement and consequent exonuclease sensitivity. Degradation was greatly enhanced by bPNA treatment in the presence of exonucleases, with ENE half-life plunging to 6 min from >24 h. RNA digestion kinetics could clearly distinguish between bPNAs with similar URIL affinities, highlighting the utility of functional assays for evaluating synthetic RNA binders. In vitro activity was mirrored by a 50% knockdown of MALAT1 expression in pancreatic cancer (PANC-1) cells upon treatment with bPNAs, consistent with intracellular digestion triggered by a similar ENE A-tail displacement mechanism. Pulldown from PANC-1 total RNA with biotinylated bPNA enriched MALAT1 > 4000× , supportive of bPNA-URIL selectivity. Together, these experiments establish the feasibility of native transcript targeting by bPNA in both in vitro and intracellular contexts. Reagents such as bPNAs may be useful tools for the investigation of transcripts stabilized by cis-acting poly(A) binding RNA elements.

The uptake of macromolecules and larger energy-rich particles into the cell is known as phagocytosis. Phagocytosed material is enzymatically degraded in membrane-bound vesicles of the endosome/lysosome system (intracellular digestion). Whereas most, if not all, cells of the animal body are equipped with the molecular apparatus for phagocytosis and intracellular digestion, a few cell types are specialized for a highly efficient mode of phagocytosis. These are the ("professional") macrophages, motile cells that seek out and eliminate pathogenic invaders or damaged cells. Macrophages form the backbone of the innate immune system. Developmentally, they derive from specialized compartments within the embryonic mesoderm and early vasculature as part of the process of hematopoiesis. Intensive research has revealed in detail molecular and cellular mechanisms of phagocytosis and intracellular digestion in macrophages. In contrast, little is known about a second type of cell that is "professionally" involved in phagocytosis, namely the "enteric phagocyte." Next to secretory (zymogenic) cells, enteric phagocytes form one of the two major cell types of the intestine of most invertebrate animals. Unlike vertebrates, these invertebrates only partially digest food material in the intestinal lumen. The resulting food particles are absorbed by phagocytosis or pinocytosis and digested intracellularly. In this review, we provide a brief overview of the enteric phagocytes described electron microscopically for diverse invertebrate clades, to then to compare these cells with the "canonical" phagocyte ultrastructure established for macrophages. In addition, we will review observations and speculations associated with the hypothesis that macrophages are evolutionarily derived from enteric phagocytes. This idea was already proposed in the late nineteenth century by Elias Metschnikoff who pioneered the research of phagocytosis for both macrophages and enteric phagocytes. We presume that modern approaches to better understand phagocytosis will be helped by considering the deep evolutionary relationship between the two cell types.

The molluskan digestive gland has been widely studied and its structural and ultrastructural descriptions have allowed the understanding of its several functions. Despite siphonarids are broadly distributed around the world, morphological studies on their digestive system are poorly represented. The panpulmonate limpet Siphonaria lessonii is the most abundant gastropod and the dominant herbivore in the rocky intertidal coast of Buenos Aires. The aim of this study was to describe the morphology, histology, ultrastructure, and histochemistry of the digestive gland of this gastropod as well as the cycle of activity of digestion. For that, different histochemical techniques along with light microscopy, transmission electron microscopy, and scanning electron microscopy were employed. This study revealed a complex epithelium, composed of a simple layer with five cell types. Digestive cells and vacuolated cells are responsible for intracellular digestion and energy accumulation; basophilic cells, secrete substances that would be involved in extracellular digestion; pigmented cells might have an excretory function and thin cells would correspond to undifferentiated cells. In addition, the tubules present a changing morphology according to the digestive activity that they undergo. As S. lessonii is a grazer that feeds continuously, the cycle of activity of the digestive gland seems to be daily. Fil: Landro, Sonia Maribel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Museo Argentino de Ciencias Naturales "Bernardino Rivadavia"; Argentina Fil: Teso, Silvia Valeria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Museo Argentino de Ciencias Naturales "Bernardino Rivadavia"; Argentina Fil: Arrighetti, Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Museo Argentino de Ciencias Naturales "Bernardino Rivadavia"; Argentina

Recent advances in the understanding of the functional relevance of sap of mature plant cells are reviewed. The emphasis is placed on roles of vacuoles played in the temporary storage of saccharides and organic acids, in the accumulation of water soluble products of secondary metabolism and in the intracellular digestion of protein. Contents Summary 1 I. Introduction 1 II. Functions of vacuoles 2 III. Vacuoles as pools of saccharides 3 IV. Organic acids 7 V. (Potentially) toxic cell saps 9 VI. Pools of protein 14 VII. Digestive cell saps 15 VIII. Tonoplast, cell sap and cytoplasm 18 IX. Cellular homeostasis 19 Acknowledgement 20 References 20.

Made available in DSpace on 2021-06-25T12:26:17Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-07-13. Added 1 bitstream(s) on 2021-07-15T15:06:08Z : No. of bitstreams: 1 S0104-64972020000100222.pdf: 14167506 bytes, checksum: 4877f38654a67aec864f7c1b58c1e4bb (MD5) Pro-Reitoria de Pesquisa, Sao Paulo State University The purpose of this study was to describe the structure of the midgut (hepatopancreas and intestine) in the endemic species, Macrobrachium carcinus. Thirty specimens were collected, and the midgut was fixed in Bouin's solution for histological and histochemical analyzes by light microscopy. The hepatopancreas consists of two lobes that connect to the end of the stomach by primary ducts, which originate secondary tubules or hepatopancreatic ducts, that branch into hepatopancreatic tubules. The hepatopancreatic duct presents a columnar epithelium composed of R- and F- cells with evident brush borders for absorption and storage. The hepatopancreatic tubule is lined by epithelium with five cell types (E, F, R, B, and M). The distal region presents all cell types, with a predominance of E-cells that correspond to epithelial renewal. The middle region presents F- and B- cells, characteristic of extracellular and intracellular digestion. The proximal region, with B- and R- cells, performs the final digestion, storage, and extrusion of the cells with waste material. The intestine is lined by a single cell type with an evident brush border, suggesting luminal absorption. This cellular arrangement along the length of the midgut proposes distinct morpho-functional characteristics of digestion, absorption, and storage in this species. Sao Paulo State Univ, Biol Sci Dept, Sch Sci, UNESP, Campus Bauru, Bauru, SP, Brazil Sao Paulo State Univ, Morphol Dept, Inst Biosci, UNESP, Campus Botucatu, Botucatu, SP, Brazil Univ Fed Rio Grande do Norte, Technol Ctr, Agr Coll Jundiai, Natal, RN, Brazil Sao Paulo State Univ, UNESP, Aquaculture Ctr, Sao Paulo, SP, Brazil Sao Paulo State Univ, Biol Sci Dept, Sch Sci, UNESP, Campus Bauru, Bauru, SP, Brazil Sao Paulo State Univ, Morphol Dept, Inst Biosci, UNESP, Campus Botucatu, Botucatu, SP, Brazil Sao Paulo State Univ, UNESP, Aquaculture Ctr, Sao Paulo, SP, Brazil Pro-Reitoria de Pesquisa, Sao Paulo State University: 45231

The study of digestive physiology is a crucial aspect enabling commercial aquaculture of Octopus vulgaris type II. Knowing the processes of digestion, absorption, and assimilation of nutrients and factors that modulate the digestive environment is fundamental for designing adequate diets based on the digestive capacity of the animals. In the present study, we characterized the activity of total proteases in the gastric juice and digestive gland in adult O. vulgaris type II both fasted and during the postprandial period, and evaluated their digestive dynamics. In vitro experiments showed this species had predominantly acidic digestion, with maximal proteolytic activity at pH 3–4 and temperatures of 50 °C and 70 °C. These results confirmed the carnivorous feeding habit and high adaptive capacity of digestive enzymes in this species. The digestion time was 400 min at 20 °C when the digestive tract was ready to receive its next meal. Digestion occurred in two stages: the first, at the beginning of digestion, with rapid passage of soluble nutrients along the digestive tract; the second, which was slower, and involved the digestion, absorption, and assimilation of more complex nutrients (proteins and lipids). The ingestion peak occurred at 200 min after feeding when proteolytic activity was maximal. This species uses proteins as its primary source of energy and mobilizes proteins before lipids during intracellular digestion, which is a different digestion pattern to other tropical and subtropical-temperate octopus species. This new information contributes to a better understanding of the digestive physiology of O. vulgaris type II and is essential for formulating specific commercial diets for its farming. Keywords: Cephalopod, Protease, Gastric juice, Digestive physiology, Aquaculture

Background Despite widespread interest in solar-powered sea slugs (Sacoglossa: Gastropoda), relatively little is know about how they actually perform functional kleptoplasty. Sister-taxa Elysia timida and E. cornigera provide an ideal model system for investigating this phenomenon, since they feed on the same algal genus and only E. timida is capable of long-term kleptoplasty. Recent research has explored factors regarding functional kleptoplasty in E. timida, including their starvation longevity, digestive activity, autophagal response and photosynthetic efficiency under two different temperature conditions (18 °C and 21 °C). These studies revealed the trends E. timida displays regarding each factor during starvation as well as influences temperature has on some aspects of functional kleptoplasty. This study examines E. cornigera regarding each of these factors in an attempt to elucidate differences between each species that could explain their differing kleptoplastic abilities. Since both species naturally occur in 25 °C seawater (E. timida peak summer temperature, E. cornigera low winter temperature), each species was acclimatized to 25 °C to facilitate comparison and determine if these species exhibit physiological differences to starvation when under the same environmental conditions. Results When comparing the different E. timida temperature treatments, it becomes clear that increased temperatures compromise E. timida’s kleptoplastic abilities. Specimens acclimatized to 25 °C revealed shorter starvation longevities surviving an average 42.4 days compared to the 95.9 day average observed in specimens exposed to 18 °C. Each temperature treatment displayed a significantly different decrease throughout the starvation period in both, the rate of photosynthetic efficiency and in the decreasing functional kleptoplast abundance. Lysosomal abundances are assessed here as indicators of different aspects of metabolic activity, which could be correlated to temperature. E. cornigera, also acclimatized to 25 °C did not display significantly similar patterns as any of the E. timida temperature treatments, having fewer incorporated kleptoplasts, a higher lysosomal response to starvation, a faster decrease in photosynthetic efficiency and a lower starvation longevity. Conclusions These results confirm that each species has different physiological reactions to starvation and kleptoplast retention, even under the same conditions. While temperature affects aspects of functional kleptoplasty, it is likely not responsible for the differences in kleptoplastic abilities seen in these species.

Black snook, Centropomus nigrescens, have been identified as a promising candidate for aquaculture although, like many of the Centropomid species, high mortality associated with early larval stages presents a significant bottleneck to their commercialization. The digestive capacity of black snook larvae throughout the first 37 d after hatch (d.a.h.) was evaluated by quantifying digestive enzyme activities using biochemical techniques. Results showed that black snook larvae have alkaline proteases at hatching, which are known to be important during the first days of feeding for digestion. Toward the end of the study, acid proteases concentration increased (37 d.a.h.). Enzymes for lipid digestion, pancreatic lipase and bile salt-activated lipase, were already present in the larvae before exogenous feeding commenced, and their activity increased with age and growth (length). Intracellular digestion, measured as the activity of leucine-alanine peptidase, was high early on (5 d.a.h.) and decreased as development progressed (next 32 d). In contrast, alkaline phosphatase activity was lowest at first feeding and subsequently increased with age. Overall patterns in enzyme activity suggest the possibility of live feed weaning before 32 d.a.h. if artificial diets can be properly balanced.

The purposes of this study are to characterize magneto-endosymbiont (ME) labeling of mammalian cells and to discern the subcellular fate of these living contrast agents. MEs are novel magnetic resonance imaging (MRI) contrast agents that are being used for cell tracking studies. Understanding the fate of MEs in host cells is valuable for designing in vivo cell tracking experiments. The ME’s surface epitopes, contrast-producing paramagnetic magnetosomal iron, and genome were studied using immunocytochemistry (ICC), Fe and MRI contrast measurements, and quantitative polymerase chain reaction (qPCR), respectively. These assays, coupled with other common assays, enabled validation of ME cell labeling and dissection of ME subcellular processing. The assays mentioned above provide qualitative and quantitative assessments of cell labeling, the subcellular localization and the fate of MEs. ICC results, with an ME-specific antibody, qualitatively shows homogenous labeling with MEs. The ferrozine assay shows that MEs have an average of 7 fg Fe/ME, ∼30 % of which contributes to MRI contrast and ME-labeled MDA-MB-231 (MDA-231) cells generally have 2.4 pg Fe/cell, implying ∼350 MEs/cell. Adjusting the concentration of Fe in the ME growth media reduces the concentration of non-MRI contrast-producing Fe. Results from the qPCR assay, which quantifies ME genomes in labeled cells, shows that processing of MEs begins within 24 h in MDA-231 cells. ICC results suggest this intracellular digestion of MEs occurs by the lysosomal degradation pathway. MEs coated with listeriolysin O (LLO) are able to escape the primary phagosome, but subsequently co-localize with LC3, an autophagy-associated molecule, and are processed for digestion. In embryos, where autophagy is transiently suppressed, MEs show an increased capacity for survival and even replication. Finally, transmission electron microscopy (TEM) of ME-labeled MDA-231 cells confirms that the magnetosomes (the MRI contrast-producing particles) remain intact and enable in vivo cell tracking. MEs are used to label mammalian cells for the purpose of cell tracking in vivo, with MRI. Various assays described herein (ICC, ferrozine, and qPCR) allow qualitative and quantitative assessments of labeling efficiency and provide a detailed understanding of subcellular processing of MEs. In some cell types, MEs are digested, but the MRI-producing particles remain. Coating with LLO allows MEs to escape the primary phagosome, enhances retention slightly, and confirms that MEs are ultimately processed by autophagy. Numerous intracellular bacteria and all endosymbiotically derived organelles have evolved molecular mechanisms to avoid intracellular clearance, and identification of the specific processes involved in ME clearance provides a framework on which to develop MEs with enhanced retention in mammalian cells.

Intracellular pH values are some of the most important factors that govern biological processes and the acid-base homeostasis in cells, body fluids and organs sustains the normal operations of the body. Subcellular organelles including the acidic lysosomes and the alkalescent mitochondria undergo various processes such as intracellular digestion, ATP production and apoptosis. Due to their precise imaging capabilities, fluorescent probes have attracted great attention for the illustration of pH modulated processes. Furthermore, based on the unique acidic extracellular environment of acidic lysosomes, fluorescent probes can specifically be activated in cancer cells or tumors. In this review, recently reported lysosome and mitochondria specific pH imaging probes as well as pH-activatable cancer cell-targetable probes have been discussed.

The guts of neonatal mammals and stomachless fish have a limited capacity for luminal protein digestion, which allows oral acquisition of antibodies and antigens. However, how dietary protein is absorbed during critical developmental stages when the gut is still immature is unknown. Here, we show that specialized intestinal cells, which we call lysosome-rich enterocytes (LREs), internalize dietary protein via receptor-mediated and fluid-phase endocytosis for intracellular digestion and trans-cellular transport. In LREs, we identify a conserved endocytic machinery, composed of the scavenger receptor complex Cubilin/Amnionless and Dab2, that is required for protein uptake by LREs and for growth and survival of larval zebrafish. Moreover, impairing LRE function in suckling mice, via conditional deletion of Dab2, leads to stunted growth and severe protein malnutrition reminiscent of kwashiorkor, a devastating human malnutrition syndrome. These findings identify digestive functions and conserved molecular mechanisms in LREs that are crucial for vertebrate growth and survival.

Digestive systems and extracellular digestion are key animal features, but their emergence during early animal evolution is currently poorly understood. As the last common ancestor of non-bilaterian animal groups (sponges, ctenophores, placozoans and cnidarians) dates back to the beginning of animal life, their study and comparison provides important insights into the early evolution of digestive systems and functions. Here, I have compiled an overview of the development and cell biology of digestive tissues in non-bilaterian animals. I will highlight the fundamental differences between extracellular and intracellular digestive processes, and how these are distributed among animals. Cnidarians (e.g. sea anemones, corals, jellyfish), the phylogenetic outgroup of bilaterians (e.g. vertebrates, flies, annelids), occupy a key position to reconstruct the evolution of bilaterian gut evolution. A major focus will therefore lie on the development and cell biology of digestive tissues in cnidarians, especially sea anemones, and how they compare to bilaterian gut tissues. In that context, I will also review how a recent study on the gastrula fate map of the sea anemone Nematostella vectensis challenges our long-standing conceptions on the evolution of cnidarian and bilaterian germ layers and guts. publishedVersion

Solar-powered sea slugs (Sacoglossa: Gastropoda) have long captured the attention of laymen and scientists alike due to their remarkable ability to steal functional chloroplasts from their algal food, enslaving them to withstand long starvation periods. Recently, a wealth of data has shed insight into this remarkable relationship; however, the cellular mechanisms governing this process are still completely unknown. This study explores these mechanisms, providing insight into the chloroplast retention and delayed digestion, occurring within the slug’s digestive gland. We examine the relationships between functional chloroplast and lysosome abundances during starvation, in live material, for the long-term retaining species Elysia timida, the ambiguous long/short-term retaining Elysia viridis, and the short-term retaining Thuridilla hopei, to elucidate digestive differences that contribute to the development of functional kleptoplasty. Functional chloroplast and lysosome abundance are measured using chlorophyll a autofluorescence and the pH-dependent stain acridine orange. In each species, the number of chloroplasts and lysosomes is indirectly proportional, with the plastid density decreasing when starvation begins. We also present a new FIJI/Image J Plugin, the 3D—Accounting and Measuring Plugin, 3D-AMP, which enables the reliable analysis of large image sets.

Made available in DSpace on 2018-12-11T16:58:39Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-01-20 The morphological development of the digestive system of Hemisorubim platyrhynchos was studied from the day of hatching until 21. days post-hatching (DPH) using histology, histochemistry and scanning electron microscopy to augment the available knowledge regarding the organogenesis of the digestive system of this carnivorous Neotropical fish. The development of the digestive system was divided into four major stages. Stage I (endotrophic period) starts with hatching and ends with the mouth opening at 2 DPH. The digestive tract originated as a straight undifferentiated tube and ended as an esophagus with goblet cells, an incipient stomach and an intestine divided into the anterior, middle, posterior and rectum. Stage II (endo-exotrophic period) is from the onset of feeding to exhaustion of the yolk at 4 DPH. Stage III is the period in which the larvae rely exclusively on exogenous feeding but still have no functional stomach. Stage IV is an exotrophic period marked by the appearance of gastric glands at 15 DPH. At 20 DPH, the saccular stomach can be observed. Fish growth was largely variable over the time period studied, and the variability was predominant between the period in which the yolk was present and after its exhaustion. The mixed feeding period is short, and the larvae subsequently survive solely by exogenous feeding without a functional stomach for 15. days. During this period, the primary site of digestion is the anterior intestine, which presents with a saccular shape. The accessory glands, liver and pancreas were differentiated at 2 DPH and thus contributed to extracellular digestion. Also observed in the intestine were supranuclear inclusions that could promote intracellular digestion. The rectal columnar epithelium showed scarce goblet cells but had apical mucosubstances that were involved in fecal transit, epithelial protection and in the final absorption of substances. Gastric glands appeared at 15 DPH and, until this period, the larvae should receive live feed and after 15 DPH may be weaned. H. platyrhynchos farmers should also be alert to differences in the size of the fish because this species presents cannibalism and needs to be separated into homogeneous batches. Department of Biological Sciences Faculty of Sciences and Aquaculture Center São Paulo State University-UNESP Institute of Biosciences Letters and Exact Sciences São Paulo State University-UNESP Hydrobiology and Aquaculture Station of Sao Paulo Energetic Company-CESP Department of Biological Sciences Faculty of Sciences and Aquaculture Center São Paulo State University-UNESP Institute of Biosciences Letters and Exact Sciences São Paulo State University-UNESP

Culture of octopuses is still in an experimental stage due to a lack of knowledge about their digestive physiology. This study aimed to determine the temporality of nutrient digestion, absorption and assimilation in 2 species of octopus: the tropical Octopus maya (habitat temperature 22 to 26°C) and the subtropical-temperate O. mimus (15 to 21°C). In both species, the chyme volume displayed 2 peaks over time along the digestive tract: one at the beginning of digestion and a second at the end of the digestion process. These phases were observed in gastric juice (GJ) enzymes, in the flow of nutrients in the digestive gland (DG), in the appearance of metabolites in the hemo lymph, and in muscle protein and glycogen levels. The soluble nutrient flow throughout the digestive tract into the DG was similar in both species. How ever, intracellular digestion was different in O. mimus compared to O. maya; lipids were mobilized faster than proteins in O. mimus, whereas proteins were mobilized faster than lipids in O. maya. This suggests that at low temperatures, the mobilization of lipids could be a biological priority due to their role in membrane protection. In addition, O. mimus displayed more digestive enzyme activity in the GJ and DG compared with O. maya, suggesting that adaptive differences may be related to environmental temperatures.

Vacuole of eukaryotic cells, beyond intracellular digestion plays additional roles such as storage of nutrients that provide favorable conditions for bacterial survival. In this study, occurrence of H. pylori inside the vacuole of Candida yeast was studied and the role of vacuolating cytotoxin A (VacA) in constructing the vacuole was discussed. One gastric Candida yeast was used for Live/Dead stain and fluorescence in situ hybridization (FISH) with universal bacterial probe. Yeast total DNA was used for amplification of full-length bacterial 16S rDNA as well as H. pylori-specific 16S rDNA and vacA alleles. Vacuoles were isolated from yeast cells and stained with fluorescent yeast vacuole membrane marker MDY-64. DNA extracted from vacuoles was used for amplification of H. pylori-specific 16S rDNA. Fluorescent microscopy showed occurrence of viable bacteria inside the vacuole of intact Candida yeast cells. FISH showed intracellular bacteria as fluorescent spots inside the vacuole of mother and daughter yeast cells, suggesting bacterial transmission to next generations of yeast. Sequencing of amplified products of bacterial 16S rDNA and amplification of H. pylori 16S rDNA and vacA confirmed the identity of intracellular bacteria as H. pylori. Isolated vacuoles were stained with membrane-specific marker and H. pylori 16S rDNA was amplified from their DNA content. Results of this study suggest yeast vacuole as a specialized niche for H. pylori. It appears that sequestration inside the vacuole may enhance bacterial survival.

The two-spotted spider mite (TSSM), Tetranychus urticae Koch (Acari: Tetranychidae), is one of the most polyphagous herbivores, feeding on more than 1,100 plant species. Its wide host range suggests that TSSM has an extraordinary ability to modulate its digestive and xenobiotic physiology. The analysis of the TSSM genome revealed the expansion of gene families that encode proteins involved in digestion and detoxification, many of which were associated with mite responses to host shifts. The majority of plant defense compounds that directly impact mite fitness are ingested. They interface mite compounds aimed at counteracting their effect in the gut. Despite several detailed ultrastructural studies, our knowledge of the TSSM digestive tract that is needed to support the functional analysis of digestive and detoxification physiology is lacking. Here, using a variety of histological and microscopy techniques, and a diversity of tracer dyes, we describe the organization and properties of the TSSM alimentary system. We define the cellular nature of floating vesicles in the midgut lumen that are proposed to be the site of intracellular digestion of plant macromolecules. In addition, by following the TSSM's ability to intake compounds of defined sizes, we determine a cut off size for the ingestible particles. Moreover, we demonstrate the existence of a distinct filtering function between midgut compartments which enables separation of molecules by size. Furthermore, we broadly define the spatial distribution of the expression domains of genes involved in digestion and detoxification. Finally, we discuss the relative simplicity of the spider mite digestive system in the context of mite's digestive and xenobiotic physiology, and consequences it has on the effectiveness of plant defenses.

A review of acarine gut physiology based on published narratives dispersed over the historical international literature is given. Then, in an experimental study of the free-living predatory soil mite Pergamasus longicornis (Berlese), quantitative micro-anatomical changes in the gut epithelium are critically assessed from a temporal series of histological sections during and after feeding on larval dipteran prey. An argued functional synthesis based upon comparative kinetics is offered for verification in other mesostigmatids. Mid- and hind-gut epithelia cell types interconvert in a rational way dependent upon the physical consequences of ingestion, absorption and egestion. The fasted transitional pseudo-stratified epithelium rapidly becomes first squamous on prey ingestion (by stretching), then columnar during digestion before confirmed partial disintegration (gut 'lumenation') during egestion back to a pseudo-stratified state. Exponential processes within the mid- and endodermic hind-gut exhibit 'stiff' dynamics. Cells expand rapidly ([Formula: see text] 22.9-49.5 min) and vacuolate quickly ([Formula: see text] 1.1 h). Cells shrink very slowly ([Formula: see text] 4.9 days) and devacuolate gently ([Formula: see text] 1.0-1.7 days). Egestive cellular degeneration has an initial [Formula: see text] 7.7 h. Digestion appears to be triggered by maximum gut expansion-estimated at 10 min post start of feeding. Synchrony with changes in gut lumen contents suggests common changes in physiological function over time for the cells as a whole tightly-coupled epithelium. Distinct in architecture as a tissue over time the various constituent cell types appear functionally the same. Functional phases are: early fluid transportation (0-1 h) and extracellular activity (10-90 min); through rising food absorption (10 min to [Formula: see text] day); to slow intracellular meal processing and degenerative egestive waste material production (1 to [Formula: see text] days) much as in ticks. The same epithelium is both absorptive and degenerative in role. The switch in predominant physiology begins 4 h after the start of feeding. Two separate pulses of clavate cells appear to be a mechanism to facilitate transport by increasing epithelial surface area in contact with the lumen. Free-floating cells may augment early extracellular lumenal digestion. Possible evidence for salivary enzyme alkaline-related extra-corporeal digestion was found. Giant mycetome-like cells were found embedded in the mid-gut wall. Anteriorly, the mid-gut behaves like a temporally expendable food processing tissue and minor long-term resistive store. Posteriorly the mid-gut behaves like a major assimilative/catabolic tissue and 'last-out' food depot (i.e., a 'hepatopancreas' function) allowing the mite to resist starvation for up to 3.5 weeks after a single meal. A 'conveyor-belt' wave of physiology (i.e., feeding and digestion, then egestion and excretion) sweeps posteriorly but not necessarily pygidially over time. Assimilation efficiency is estimated at 82%. The total feeding cycle time histologically from a single meal allowing for the bulk of intracellular digestion and egestive release is not 52.5 h but of the order of 6 days ([Formula: see text] total gut emptyings per day), plus typically a further 3 days for subsequent excretion to occur. Final complete gut system clearance in this cryptozooid may take much longer ([Formula: see text] days). A common physiology across the anactinotrichid acarines is proposed. A look to the future of this field is included.

The digestive methods employed by amphioxus (Branchiostoma)—both intracellular phagocytic digestion and extracellular digestion—have been discussed since 1937. Recent studies also show that epithelial cells lining the Branchiostoma digestive tract can express many immune genes. Here, in Branchiostoma belcheri, using a special tissue fixation method, we show that some epithelial cells, especially those lining the large diverticulum protruding from the gut tube, phagocytize food particles directly, and Branchiostoma can rely on this kind of phagocytic intracellular digestion to obtain energy throughout all stages of its life. Gene expression profiles suggest that diverticulum epithelial cells have functional features of both digestive cells and phagocytes. In starved Branchiostoma, these cells accumulate endogenous digestive and hydrolytic enzymes, whereas, when sated, they express many kinds of immune genes in response to stimulation by phagocytized food particles. We also found that the distal hindgut epithelium can phagocytize food particles, but not as many. These results illustrate phagocytic intercellular digestion in Branchiostoma, explain why Branchiostoma digestive tract epithelial cells express typical immune genes and suggest that the main physiological function of the Branchiostoma diverticulum is different from that of the vertebrate liver.

Cysteine cathepsins are widely spread on living organisms associated to protein degradation in lysosomes, but some groups of Arthropoda (Heteroptera, Coleoptera, Crustacea and Acari) present these enzymes related to digestion of the meal proteins. Although spiders combine a mechanism of extra-oral with intracellular digestion, the sporadic studies on this subject were mainly concerned with the digestive fluid (DF) analysis. Thus, a more complete scenario of the digestive process in spiders is still lacking in the literature. In this paper we describe the identification and characterization of cysteine cathepsins in the midgut diverticula (MD) and DF of the spider Nephilengys cruentata by using enzymological assays. Furthermore, qualitative and quantitative data from transcriptomic followed by proteomic experiments were used together with biochemical assays for results interpretation. Five cathepsins L, one cathepsin F and one cathepsin B were identified by mass spectrometry, with cathepsins L1 (NcCTSL1) and 2 (NcCTSL2) as the most abundant enzymes. The native cysteine cathepsins presented acidic characteristics such as pH optima of 5.5, pH stability in acidic range and zymogen conversion to the mature form after in vitro acidification. NcCTSL1 seems to be a lysosomal enzyme with its recombinant form displaying acidic characteristics as the native ones and being inhibited by pepstatin. Evolutionarily, arachnid cathepsin L may have acquired different roles but its use for digestion is a common feature to studied taxa. Now a more elucidative picture of the digestive process in spiders can be depicted, with trypsins and astacins acting extra-orally under alkaline conditions whereas cysteine cathepsins will act in an acidic environment, likely in the digestive vacuoles or lysosome-like vesicles.

Background Despite widespread interest in solar-powered sea slugs (Sacoglossa: Gastropoda), relatively little is know about how they actually perform functional kleptoplasty. Sister-taxa Elysia timida and E. cornigera provide an ideal model system for investigating this phenomenon, since they feed on the same algal genus and only E. timida is capable of long-term kleptoplasty. Recent research has explored factors regarding functional kleptoplasty in E. timida, including their starvation longevity, digestive activity, autophagal response and photosynthetic efficiency under two different temperature conditions (18 °C and 21 °C). These studies revealed the trends E. timida displays regarding each factor during starvation as well as influences temperature has on some aspects of functional kleptoplasty. This study examines E. cornigera regarding each of these factors in an attempt to elucidate differences between each species that could explain their differing kleptoplastic abilities. Since both species naturally occur in 25 °C seawater (E. timida peak summer temperature, E. cornigera low winter temperature), each species was acclimatized to 25 °C to facilitate comparison and determine if these species exhibit physiological differences to starvation when under the same environmental conditions. Results When comparing the different E. timida temperature treatments, it becomes clear that increased temperatures compromise E. timida’s kleptoplastic abilities. Specimens acclimatized to 25 °C revealed shorter starvation longevities surviving an average 42.4 days compared to the 95.9 day average observed in specimens exposed to 18 °C. Each temperature treatment displayed a significantly different decrease throughout the starvation period in both, the rate of photosynthetic efficiency and in the decreasing functional kleptoplast abundance. Lysosomal abundances are assessed here as indicators of different aspects of metabolic activity, which could be correlated to temperature. E. cornigera, also acclimatized to 25 °C did not display significantly similar patterns as any of the E. timida temperature treatments, having fewer incorporated kleptoplasts, a higher lysosomal response to starvation, a faster decrease in photosynthetic efficiency and a lower starvation longevity. Conclusions These results confirm that each species has different physiological reactions to starvation and kleptoplast retention, even under the same conditions. While temperature affects aspects of functional kleptoplasty, it is likely not responsible for the differences in kleptoplastic abilities seen in these species. Electronic supplementary material The online version of this article (10.1186/s12983-018-0264-y) contains supplementary material, which is available to authorized users.

Mid- and hind-gut lumenal changes are described in the free-living predatory soil mite Pergamasus longicornis (Berlese) from a time series of histological sections scored during and after feeding on fly larval prey. Three distinct types of tangible material are found in the lumen. Bayesian estimation of the change points in the states of the gut lumenal contents over time is made using a time-homogenous first order Markov model. Exponential processes within the gut exhibit ‘stiff’ dynamics. A lumen is present throughout the midgut from 5 min after the start of feeding as the gut rapidly expands. It peaks at about 21.5 h–1.5 days and persists post-feeding (even when the gut is contracted) up until fasting/starvation commences 10 days post start of feeding. The disappearance of the lumen commences 144 h after the start of feeding. Complete disappearance of the gut lumen may take 5–9 weeks from feeding commencing. Clear watery prey material arrives up to 10 min from the start of feeding, driving gut lumen expansion. Intracellular digestion triggered by maximum gut expansion is indicated. Detectable granular prey material appears in the lumen during the concentrative phase of coxal droplet production and, despite a noticeable collapse around 12 h, lasts in part for 52.5 h. Posterior midgut regions differ slightly from anterior regions in their main prey food dynamics being somewhat faster in processing yet being slightly delayed. Posterior regions are confirmed as Last-In-Last-Out depots, anterior regions confirmed as First-In-First-Out conveyor belt processes. Evidence for differential lability of prey fractions is found. A scheme is presented of granular imbibed prey material being first initially rapidly absorbed (\documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$t_{\frac{1}{2}}$$\end{document}t12 = 23 min), and also being quickly partly converted to globular material extra-corporeally/extracellularly (\documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$t_{\frac{1}{2}}$$\end{document}t12 = 36 min)—which then rapidly disappears (\documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$t_{\frac{1}{2}}$$\end{document}t12 = 1.1 h, from a peak around 4 h). This is then followed by slow intracellular digestion (\documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$t_{\frac{1}{2}}$$\end{document}t12 = 6.9 h) of the resultant resistant prey residue matching the slow rate of appearance of opaque pre-excretory egestive refractive grains (overall \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$t_{\frac{1}{2}}$$\end{document}t12 = 4.5 days). The latter confirmed latent ‘catabolic fraction’ (along with Malpighian tubule produced guanine crystals) drives rectal vesicle expansion as ‘faeces’ during the later phases of gut emptying/contraction. Catabolic half-lives are of the order of 6.3–7.8 h. Membraneous material is only present in the lumen of the gut in starving mites. No obvious peritrophic membrane was observed. The total feeding cycle time may be slightly over 52.5 h. Full clearance in the gut system of a single meal including egestive and excretory products may take up to 3 weeks. Independent corroborative photographs are included and with posterior predictive densities confirm the physiological sequence of ingestion/digestion, egestion, excretion, defecation, together with their timings. Visually dark midguts almost certainly indicate egestive refractive grains (xanthine?) production. Nomograms to diagnose the feeding state of P. longicornis in field samples are presented and show that the timing of these four phases in the wild could be inferred by scoring 10–12 mites out of a sample of 20. Suggestions to critically confirm or refute the conclusions are included.

Plague is a flea-borne rodent-associated zoonotic disease caused by Yersinia pestis . The disease is characterized by epizootics with high rodent mortalities, punctuated by interepizootic periods when the bacterium persists in an unknown reservoir. This study investigates the interaction between Y. pestis and the ubiquitous soil free-living amoeba (FLA) Acanthamoeba castellanii to assess if the bacterium can survive within soil amoebae and whether intracellular mechanisms are conserved between infection of mammalian macrophages and soil amoebae. The results demonstrate that during coculture with amoebae, representative Y. pestis strains of epidemic biovars Medievalis, Orientalis, and Antiqua are phagocytized and able to survive within amoebae for at least 5 days. Key Y. pestis determinants of the intracellular interaction of Y. pestis and phagocytic macrophages, PhoP and the type three secretion system (T3SS), were then tested for their roles in the Y. pestis -amoeba interaction. Consistent with a requirement for the PhoP transcriptional activator in the intracellular survival of Y. pestis in macrophages, a PhoP mutant is unable to survive when cocultured with amoebae. Additionally, induction of the T3SS blocks phagocytic uptake of Y. pestis by amoebae, similar to that which occurs during macrophage infection. Electron microscopy revealed that in A. castellanii , Y. pestis resides intact within spacious vacuoles which were characterized using lysosomal trackers as being separated from the lysosomal compartment. This evidence for prolonged survival and subversion of intracellular digestion of Y. pestis within FLA suggests that protozoa may serve as a protective soil reservoir for Y. pestis . IMPORTANCE Yersinia pestis is a reemerging flea-borne zoonotic disease. Sylvatic plague cycles are characterized by an epizootic period during which the disease spreads rapidly, causing high rodent mortality, and an interepizootic period when the bacterium quiescently persists in an unknown reservoir. An understanding of the ecology of Y. pestis in the context of its persistence in the environment and its reactivation to initiate a new epizootic cycle is key to implementing novel surveillance strategies to more effectively predict and prevent new disease outbreaks. Here, we demonstrate prolonged survival and subversion of intracellular digestion of Y. pestis within a soil free-living amoeba. This suggests the potential role for protozoa as a protective soil reservoir for Y. pestis , which may help explain the recrudescence of plague epizootics.

Larval fish have a relatively poorly developed immune system and hence they primarily rely on non-specific immune responses produced in the skin and gut. This study aims to illustrate the fine structure of gut development in common dentex (Dentex dentex) larvae and determine the initiation of pinocytic activity, which is an important early non-specific immune response in fish intestines, during various live-food feeding stages (yeast enriched rotifer, Artemia nauplii and metanauplii) from hatching (D0) to 25 days after hatching (D25) by using transmission electron microscopy (TEM). A total of 54 larvae were sampled and examined with electron microscopy. The results show that hind-gut intestine enterocytes have morphological features which suggest they play a role in absorption and intracellular digestion of nutrients in fish. The presence of vigorous pinocytic activity in the Artemia feeding stage between 17-25 days after hatching is evidence for the development of a non-specific immune system. In the present study, the ultrastructural gut development of cultured common dentex was investigated for the first time and the data reported in this study may provide additional fundamental knowledge for improving larval rearing success in common dentex culture.

The pike-perch (Sander lucioperca L) at the age of 18 days were fed for 21 days with three different diets: Aglo Norse (AN), casein-gelatin (Cas), cod meal with gelatin (Mac) and nauplius Artemia salina (Art—control diet). On the last day of the experiment, fish fed Art and AN diets had the statistically significant highest body mass, length, and survival. On the last day of the experiment, pikes fed Cas diet had the lowest folds of the anterior and posterior intestine. In the anterior intestine of fish fed Cas diet was found a statistically significant increase in the proliferation of the enterocytes as compared with fish from other feeding groups. The fish from Mac-feeding group had the largest statistically significant total area of enterocytes and the absorption area of posterior intestine. This was probably the effect of disorders in the intracellular digestion and the stoppage in transportation of the intracellular digestion products into the circulatory system. Moreover, there was found the intensive processes of apoptosis of enterocytes in the fish fed Mac. On the last day of the experiment, fish fed Art and AN had statistically significantly more cells secreting cholecystokinin and gastrin (Gas) as compared with fish from other feeding groups. Results indicate that juvenile pike-perch can be reared and fed commercial diet AN since its 18th day after hatching. Feeding pike-perch Cas and Mac diets did not satisfy nutritional needs of fish, resulting in low survival, growth rate, and pathological changes in the intestine.

All life stages of B. chanayi (Acariformes: Cheyletidae) are characterized by occasional bloodsucking and a long period of digestion. No newly engorged mites were found during the period of their host birds' migration. The fine structure of the digestive tract of a blood-feeding acariform mite is described for the first time. The anterior midgut (AMG) is a place of blood digestion, while the posterior midgut (PMG) is involved in nitrogen metabolism forming guanine crystals as the main end-product. The AMG epithelium consists of digestive cells that probably arise from mitotically active basal cells with high synthesizing activity. As observed in ticks, blood digestion is accompanied by the formation of huge endosomes that serve as places of storage and sorting of ingested material. Digestive cells show different types of endocytotic activity as well as various late endosomes, which implies different subcellular pathways for different blood components. In both midgut regions, elimination of the excretory material occurs by apocrine secretion or by discharging of apical cell fragments (loaded with lysosomes) into the gut lumen. The formation of guanine granules occurs inside the lysosomes of PMG epithelial cells thus having much in common with intracellular digestion. Peculiarities of intracellular blood digestion were analyzed according to the modern hypothesis of endocytosis and compared to what is known in ticks.

Assessment of the potential allergenicity (IgE-inducing properties) of novel proteins is an important challenge in the overall safety assessment of foods. Resistance to digestion with pepsin is commonly measured to characterize allergenicity, although the association is not absolute. We have previously shown that specific IgE antibody production induced by systemic [intraperitoneal (i.p.)] exposure of BALB/c strain mice to a range of proteins correlates with allergenic potential for known allergens. The purpose of the present study was to explore further the utility of these approaches using the food allergen, actinidin. Recently, kiwifruit has become an important allergenic foodstuff, coincident with its increased consumption, particularly as a weaning food. The ability of the kiwifruit allergen actinidin to stimulate antibody responses has been compared with the reference allergen ovalbumin, and with the non-allergen bovine haemoglobin. Haemoglobin was rapidly digested by pepsin whereas actinidin was resistant unless subjected to prior chemical reduction (reflecting intracellular digestion conditions). Haemoglobin stimulated detectable IgG antibody production at relatively high doses (10%), but failed to provoke detectable IgE. In contrast, actinidin was both immunogenic and allergenic at relatively low doses (0.25% to 1%). Vigorous IgG and IgG1 antibody and high titre IgE antibody responses were recorded, similar to those provoked by ovalbumin. Thus, actinidin displays a marked ability to provoke IgE, consistent with allergenic potential. These data provide further encouragement that in tandem with analysis of pepsin stability, the induction of IgE after systemic exposure of BALB/c strain mice provides a useful approach for the prospective identification of protein allergens.

The digestive gland of crustacean is involved in various metabolic activities, including the synthesis and secretion of digestive enzymes that begin the process of food digestion, intracellular digestion and absorption of nutrients, storage of reserves, and disposal of waste products. It consists of two glandular lobes which extensively subdivide to form a complex of blind-ending tubules, whose size, surface area, and digestive cells are associated with intracellular digestion and the nutritional status of the organism. The aim of this paper was to study the morphology of the digestive gland in various lobster species and calculate the surface area of tubules, lumen and digestive cells (R-, F-, and B-cells) and their ratios to total tubule surface area. The similarity in ratios obtained in this study between individual lobsters suggests that the method developed in this study can be successfully applied to a range of species. This study describes a novel image processing algorithm for the automatic measurement of the hepatopancreas structure using stained cross sections of digestive gland tubules. The proposed new methodology could be used for studying the physiology and nutrient metabolism of lobsters and other crustaceans. The computer-aided analysis described in this paper is accurate for the quantitative assessment of the lobster's digestive gland structure.

Made available in DSpace on 2018-12-11T17:07:14Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-09-01 This cytochemical study investigated the acid and alkaline phosphatase of the digestive tract of Hemisorubim platyrhynchos. Acid phosphatase was detected in the lining epithelium throughout the digestive tract, whereas alkaline phosphatase was only observed in the intestine. In the esophagus, an acid phosphatase reaction occurred in the apical cytoplasm of the epithelial cells and was related to epithelial protection and freeing of superficial cells for sloughing. Similar results were also observed in epithelial cells of gastric epithelium. In the gastric glands, acid phosphatase occurred in lysosomes of the oxynticopeptic cells acting in the macromolecule degradation for use as an energy source, whereas in the vesiculotubular system, its presence could be related to secretion processes. Furthermore, acid phosphatase in the intestine occurred in microvilli and lysosomes of the enterocytes and was correlated to absorption and intracellular digestion. However, no difference was reported among the regions of the intestine. However, alkaline phosphatase reaction revealed a large number of reaction dots in the anterior intestine, with the number decreasing toward the posterior intestine. This enzyme has been related to several functions, highlighting its role in the nutrient absorption primarily in the anterior intestine but also being essential in pH regulation because this is a carnivorous species with many gastric glands with secretions that could damage the intestine. Federal University of Uberlândia – UFU Institute of Biomedical Sciences, 1720 Pará Avenue São Paulo State University – UNESP Institute of Biosciences Letter and Exact Sciences, 2265 Cristovão Colombo Street São Paulo State University – UNESP Department of Biological Sciences Faculty of Sciences, 14-01 Engenheiro Luiz Edmundo Carrijo Coube Avenue São Paulo State University – UNESP Aquaculture Center of UNESP – CAUNESP, Via de Acesso Prof. Paulo Donato Castellane, s/n São Paulo State University – UNESP Institute of Biosciences Letter and Exact Sciences, 2265 Cristovão Colombo Street São Paulo State University – UNESP Department of Biological Sciences Faculty of Sciences, 14-01 Engenheiro Luiz Edmundo Carrijo Coube Avenue São Paulo State University – UNESP Aquaculture Center of UNESP – CAUNESP, Via de Acesso Prof. Paulo Donato Castellane, s/n

In order to understand the digestive physiology of Sepia pharaonis Ehrenberg 1831 (Sepiidae) paralarvae, and to formulate appropriate feeding strategies, the activity patterns of major digestive enzymes in relation to larval development were investigated. Trypsin, leucine aminopeptidase, alkaline phosphatase, alpha-amylase, and lipase were analyzed for 25 days from hatching. Results revealed that all digestive enzymes were present even prior to exogenous feeding, and each enzyme exhibited a distinct activity pattern. Leucine aminopeptidase, an enzyme involved in intracellular digestion, was found to be dominant and highly active at early developmental stages and declined with larval maturation. Lipase activity, involved in lipid metabolism, increased in the early stages of development but declined 10 days after hatching (DAH). Secreted enzymes (trypsin and amylase) and alkaline phosphatase (an indicator of larval gut maturity) exhibited low activity prior to 10 DAH and 13 DAH, respectively. However, these enzymes became highly active in the late developmental stages. These findings suggest the gradual gut maturation of S. pharaonis based on intracellular and secreted enzyme activities. The findings also indicate that paralarvae after hatching are well-equipped with digestive enzymes necessary to digest complex food items, and that the larval gut matures 13 to 20 DAH.

Background Spiders are known for their predatory efficiency and for their high capacity of digesting relatively large prey. They do this by combining both extracorporeal and intracellular digestion. Whereas many high throughput (“-omics”) techniques focus on biomolecules in spider venom, so far this approach has not yet been applied to investigate the protein composition of spider midgut diverticula (MD) and digestive fluid (DF). Results We here report on our investigations of both MD and DF of the spider Nephilingis (Nephilengys) cruentata through the use of next generation sequencing and shotgun proteomics. This shows that the DF is composed of a variety of hydrolases including peptidases, carbohydrases, lipases and nuclease, as well as of toxins and regulatory proteins. We detect 25 astacins in the DF. Phylogenetic analysis of the corresponding transcript(s) in Arachnida suggests that astacins have acquired an unprecedented role for extracorporeal digestion in Araneae, with different orthologs used by each family. The results of a comparative study of spiders in distinct physiological conditions allow us to propose some digestion mechanisms in this interesting animal taxon. Conclusion All the high throughput data allowed the demonstration that DF is a secretion originating from the MD. We identified enzymes involved in the extracellular and intracellular phases of digestion. Besides that, data analyses show a large gene duplication event in Araneae digestive process evolution, mainly of astacin genes. We were also able to identify proteins expressed and translated in the digestive system, which until now had been exclusively associated to venom glands. Electronic supplementary material The online version of this article (doi:10.1186/s12864-016-3048-9) contains supplementary material, which is available to authorized users.